TY - JOUR
T1 - Dynamics of Translation of Single mRNA Molecules in Vivo
AU - Yan, Xiaowei
AU - Hoek, Tim A.
AU - Vale, Ronald D.
AU - Tanenbaum, Marvin E.
PY - 2016/5/5
Y1 - 2016/5/5
N2 - Regulation of mRNA translation, the process by which ribosomes decode mRNAs into polypeptides, is used to tune cellular protein levels. Currently, methods for observing the complete process of translation from single mRNAs in vivo are unavailable. Here, we report the long-term (>1 hr) imaging of single mRNAs undergoing hundreds of rounds of translation in live cells, enabling quantitative measurements of ribosome initiation, elongation, and stalling. This approach reveals a surprising heterogeneity in the translation of individual mRNAs within the same cell, including rapid and reversible transitions between a translating and non-translating state. Applying this method to the cell-cycle gene Emi1, we find strong overall repression of translation initiation by specific 5′ UTR sequences, but individual mRNA molecules in the same cell can exhibit dramatically different translational efficiencies. The ability to observe translation of single mRNA molecules in live cells provides a powerful tool to study translation regulation.
AB - Regulation of mRNA translation, the process by which ribosomes decode mRNAs into polypeptides, is used to tune cellular protein levels. Currently, methods for observing the complete process of translation from single mRNAs in vivo are unavailable. Here, we report the long-term (>1 hr) imaging of single mRNAs undergoing hundreds of rounds of translation in live cells, enabling quantitative measurements of ribosome initiation, elongation, and stalling. This approach reveals a surprising heterogeneity in the translation of individual mRNAs within the same cell, including rapid and reversible transitions between a translating and non-translating state. Applying this method to the cell-cycle gene Emi1, we find strong overall repression of translation initiation by specific 5′ UTR sequences, but individual mRNA molecules in the same cell can exhibit dramatically different translational efficiencies. The ability to observe translation of single mRNA molecules in live cells provides a powerful tool to study translation regulation.
UR - http://www.scopus.com/inward/record.url?scp=84965127033&partnerID=8YFLogxK
U2 - 10.1016/j.cell.2016.04.034
DO - 10.1016/j.cell.2016.04.034
M3 - Article
C2 - 27153498
AN - SCOPUS:84965127033
SN - 0092-8674
VL - 165
SP - 976
EP - 989
JO - Cell
JF - Cell
IS - 4
ER -